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Understanding and preventing E.Coli resistance at the abattoirs

Project Code: FOS.01.17
Completed: In Progress. Results expected in March 2022.

Project Title:

If E. coli shed by cattle is becoming resistant to antimicrobial interventions in abattoirs, how best to raise the hurdles

Researchers:

Xianqin Yang Ph.D. and Claudia Narvaez Ph.D. xianqin.yang@agr.gc.ca

Xianqin Yang Ph.D. (Agriculture Agri-Food Canada Lacombe), Claudia Narvaez Ph.D. (University of Manitoba), Tim McAllister Ph.D., (Agriculture Agri-Food Canada Lethbridge); Kim Stanford Ph.D., Tim Reuter Ph.D. (Alberta Agriculture and Forestry); Dongyan Niu Ph.D., (University of Calgary) Marcelo Dubiel, Harshita Chaudhary Ph.D., (Exigence Technologies Inc.)

Background:

Packing plants have greatly improved their control of E. coli (including Shiga toxin-producing E. coli, or STEC) on carcasses by implementing steam, hot water and organic acid interventions. They are also focusing increased attention on the proper sanitation of gloves, knives, and both fixed and moving equipment to prevent re-contamination of beef. Limited laboratory-based evidence suggests that E. coli may be surviving some of these interventions. If E. coli are evolving and adapting to these interventions, the effectiveness of current packing plant interventions may decline over time.

The cleaning agents used to wash and sanitize packing plant equipment are effective while they’re in contact with the equipment, but are eventually rinsed off, and are no longer effective. A coating of N-halamine on equipment surfaces may help to prevent this problem. With an N-halamine coating, chloride ions from bleach are attracted to the surface and stay on even after rinsing, so it will remain active until the next shift starts.

Objectives:

To Determine if E. coli from cattle are increasing in resistance to heat, acid, and sanitizers and in biofilm forming potential and the mechanisms by which E. coli persist in beef packing facilities. Researchers will identify genetic elements that confer resistance to E. coli using whole genome sequencing and determine the role and mechanisms of biofilm in the transfer of pathogens from various surfaces to meat. Researchers will also explore novel sanitization technologies to control food-borne pathogens.

What They Will Do:

The research team will characterize Top 7 STEC collected from Western Canadian slaughter plants between 2012 and 2016 (700 isolates) and feedlot cattle between 2002 through 2010 (100 isolates). These isolates will be tested for their ability to survive heat, acid, and the two most common types of sanitizers used in packing plants.

Another 750 E. coli isolates collected from hides and carcasses will be compared between a packing plant that uses complex carcass sanitation interventions and one that doesn’t; some of these isolates were also collected from equipment before and after cleaning. These isolates will also be tested for their acid, heat and sanitizer tolerance.

DNA will be compared between generic and Top 7 STEC E. coli that are resistant or susceptible to heat, acid or sanitizers, or that are able to produce biofilms. They will identify genes related to stress resistance as well as antimicrobial resistance.

E. coli strains that are known to form weak, moderate or strong biofilms will be cultured on stainless steel disks, then rinsed off and stored (damp or dry) at 4oC, 10oC and 25oC for 2, 4, 6 and 30 days. Beef samples will be placed on the disks for 5 minutes, and E. coli from the disks to the beef will be quantified.

Pieces of high density polyethylene (conveyor belt material) and stainless steel (glove material) will be coated with N-halamine. Coated and non-coated pieces will be inoculated with saline, E. coli and meat juice at 7oC (meat plant operating temperature) and 15oC (meat plant downtime temperature) and withdrawn after 0, 1, 2, 4 and 6 hours. E. coli survival and extent of cell injury will be determined. This will be repeated with pieces that have been pre-exposed to meat juice, to determine if meat juice reduces the effectiveness of the N-halamine coating. The degree of biofilm formation will be assessed. The effect of repeated N-halamine exposure on heat, acid and sanitizer tolerance of generic E. coli before and after exposure will be compared to E. coli from non-coated surfaces.

Implications:

This research will determine whether microbes are developing resistance to industry-standard heat- and acid-based food safety interventions, which is critical to assess whether additional or alternative in-plant sanitation strategies are needed to ensure the continued safety of Canadian beef for domestic and international customers.

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